Fluorescence Reader
Fluorescence Reader - Related Videos
07:51
Introduction to the Microplate Reader
0 Views •
2023
The microplate reader is a multimodal instrument that allows for a variety of experiments to be performed and measured simultaneously. Microplate readers can make absorbance, fluorescence and luminescence measurements. Multiwell plates are integral to the microplate reader and allow for many experiments to be performed at once. Regardless of the assay type, experiments on the plate reader utilize a standard curve to determine the experimental values. This curve uses samples of known...
03:01
Precise and High-Throughput Analysis of Bacterial Growth Using a Microplate Reader
0 Views •
2025
Source: Kurokawa, M., et al. Precise, High-throughput Analysis of Bacterial Growth. J. Vis. Exp. (2017)This video demonstrates the high-throughput quantification of bacterial growth using a microplate reader. It outlines the steps involved in culture preparation, optical density measurement, and analysis of growth dynamics.
03:13
Mitochondrial Calcium Uptake Assay: A Plate Reader-Based Method for Measuring Calcium Uptake in Isolated Mitochondria
0 Views •
2025
This video describes a plate reader-based assay to measure mitochondrial calcium uptake in isolated mitochondria. The assay analyzes the fluorescence signal of a calcium-sensitive fluorescent dye to study the kinetics of mitochondrial calcium uptake and calcium overload.
07:13
Characterizing Modulators of Protease-Activated Receptors with a Calcium Mobilization Assay Using a Plate Reader
0 Views •
Cited by 1 •
2024
An improved protocol for a calcium mobilization assay with endothelial cells, used to identify ligands of protease-activated receptors (PARs), has been developed. The new protocol reduces total assay time by 90-120 min and yields reproducible concentration-response curves.
02:59
Fluorescence Microplate-Based Cycloheximide Chase Assay: A Technique to Monitor the Degradation Kinetics of Fluorescent Nuclear Misfolded Proteins
0 Views •
2025
This video describes an in vitro fluorescence-based microplate assay to study the degradation kinetics of a fluorescently-labeled misfolded luciferase mutant protein expressed in the nuclei of transfected mammalian cells. The assay involves the treatment of cells expressing the fluorescent mutant protein with a translation inhibitor and a proteasome inhibitor to assess the proteasome-mediated degradation of the misfolded protein.
FAQs
The instrument illuminates fluorescent molecules at a selected excitation wavelength, which prompts them to emit light at longer wavelengths. A separate detection setting captures this emitted light rather than the incoming illumination. Using these distinct wavelength ranges allows the reader to associate the detected signal with the fluorescent label present in the sample and support quantitative analysis.
Optical filters help the instrument select the relevant emitted wavelengths before light reaches the detector. This separation is important because the reader first exposes the sample to excitation light and then measures the resulting emission. Appropriate filtering therefore supports clearer measurement of fluorescence from labeled antibodies, nucleic acid dyes, or reporter probes.
Interpretation depends on measuring the emitted light under defined conditions, including the selected excitation and emission wavelengths. The resulting signal reflects the amount or activity of the fluorescent label in the sample. Keeping those measurement conditions consistent allows signals from biological samples to be compared and used for detection or quantification.
A typical workflow places assay samples in a microplate, selects the excitation and emission settings appropriate for the fluorescent label, illuminates the wells, and records the emitted light with the detector. The resulting measurements can then be compared across samples to quantify fluorescent signals associated with antigens, antibodies, pathogens, or immune responses.
Fluorescent antibodies can provide signals associated with antigen or antibody detection, while nucleic acid dyes and reporter probes support other assay formats. In infection research, these labels help measure targets such as pathogens or host responses. The reader translates their fluorescence into data suitable for analyzing biological samples in a microplate format.
Its microplate-based format supports high-throughput analysis, allowing many samples or assay conditions to be assessed using fluorescent signals. Researchers can apply this capability to diagnostic assay development and to studies of host-pathogen interactions. Measurements may also help evaluate immune responses by detecting labeled antibodies, antigens, pathogens, or related reporter activity.